Whole mount staining of lenses for visualization of lens epithelial cell proteins

Whole mount imaging of the lens allows for high spatial resolution visualization of lens epithelial structures by using small molecule fluorescent probes. However, the visualization of specific proteins in lens epithelial cells within whole lenses remains a challenge as the capsule that surrounds th...

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Bibliographic Details
Published in:MethodsX 2021-01, Vol.8, p.101376-101376, Article 101376
Main Authors: Patel, Shaili D., Aryal, Sandeep, Mennetti, Lucas P., Parreno, Justin
Format: Article
Language:English
Subjects:
Capsule Digestion
Eye
Immunofluorescence
Lens
Method
Whole Mount immunostaining of the lens in the eye
Whole-mount
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Description
Summary:Whole mount imaging of the lens allows for high spatial resolution visualization of lens epithelial structures by using small molecule fluorescent probes. However, the visualization of specific proteins in lens epithelial cells within whole lenses remains a challenge as the capsule that surrounds the lens does not allow penetration of antibodies. Here we describe a whole mount imaging method that allows us to overcome this challenge by digesting the lens capsules of paraformaldehyde fixed lenses using collagenase. This method enables the penetration of antibodies for effective visualization of proteins in the epithelium of whole lenses.•A limitation to lens whole mount imaging is the ability to visualize specific proteins as the collagen capsule surrounding the lens impedes the penetration of antibodies•This protocol helps overcome this limitation by a light collagenase digestion of the capsule of fixed lenses prior to immunostaining•This method allows for the imaging of specific proteins in the epithelium of the whole lens tissue [Display omitted]
ISSN:2215-0161
2215-0161
DOI:10.1016/j.mex.2021.101376